Antimicrobial medical biomaterial and a method for preparing the same

ABSTRACT

An antibacterial medical biomaterial includes an acellular small intestinal submucosal matrix material, an antibacterial gel layer located on a surface of the acellular small intestinal submucosal matrix material, and an absorbable fiber layer located on a surface of the antibacterial gel layer. Sulfadiazine silver is on the surface of the acellular small intestinal submucosal matrix material and/or within the acellular small intestinal submucosal matrix material. An absorbable fiber layer to which the sulfadiazine silver is attached, wherein the content of sulfadiazine silver in the absorbable fiber is 1 wt. %˜2 wt. %. The medical biomaterial is usable as an external medicine for treating wound infections relayed by burns or wounds, and for reducing the incidence of infection by using a conventional central venous catheter with a sulfadiazine silver antibacterial coating, so that the medical biomaterial loaded with sulfadiazine silver also has antibacterial activity consistent with sulfadiazine silver.

RELATED APPLICATIONS

The present application claims priority from Chinese Application Number202010292857.0, filed Apr. 15, 2020, the disclosure of which is herebyincorporated by reference herein in its entirety.

TECHNICAL FIELD

The present invention relates to the technical field of medicalbiomaterials, and in particular to an antibacterial medical biomaterialand a method for preparing the same.

BACKGROUND

The defects of certain tissues or organs and partial or total loss offunctions caused by various diseases and traumas are one of the mainhazards to human health. Investigations and development of desirablematerials for tissue repair is always an important problem in the fieldsof medical, bioscience, and material science. Currently, in a pluralityof clinical medical biomaterials for tissue repair, an implantablemedical biomaterial is a novel biomaterial leather capable of activelyinducing tissue regeneration from a traditional non-absorbable material,and the extracellular matrix (ECM) material based on the principle oftissue engineering is a major development direction.

In the ECM material product entering the clinical application, the smallintestinal submucosal (SIS) matrix material is the most ideal softtissue repair material recognized by the academic demarcation, and theacellular SIS matrix material has a large sample amount of clinicalapplication in the fields of abdominal wall repair, burn, anus,refractory wound, shaping procedure, pelvic floor repair, tendon repair,urogenital tract repair, neurorepair, etc. However, during the clinicalapplication of the acellular SIS matrix material, the infections thatmay be generated are still problematic in the clinical problem, andtherefore, the acellular SIS matrix material itself does not haveantimicrobial and bacteriostatic capability, and it is easy to causebacteria to adhere to the surface thereof, thereby causing relevantinfections and linkage complications. Postoperative infections oftenoccur even under conditions such as full sterilization of the operatingroom and strictly aseptic surgery.

Thus, it is necessary to develop an acellular SIS matrix material withantibacterial properties.

SUMMARY

The present invention provides an antibacterial medical biomaterial anda method for preparing the same.

In one aspect, the present invention provides an antibacterial medicalbiomaterial comprising an acellular small intestinal submucosal matrixmaterial, an antibacterial gel layer located on a surface of theacellular small intestinal submucosal matrix material, and an absorbablefiber layer located on a surface of the antibacterial gel layer,sulfadiazine silver is dispersed on the surface of the acellular smallintestinal submucosal matrix material and/or within the acellular smallintestinal submucosal matrix material; the absorbable fiber layercomprises an absorbable fiber, to which the sulfadiazine silver isattached, wherein the content of sulfadiazine silver in the absorbablefiber is 1 wt. %˜2 wt. %.

Further, the absorbable fiber layer has a thickness of 0.005 mm˜0.05 mm;

Preferably, the absorbable fiber is one or more of hydroxyethylcellulose, oxidized cellulose, and carboxymethyl fiber.

Further, the content of sulfadiazine silver in the acellular smallintestinal submucosal matrix material is 2˜5 wt. %.

Further, the acellular small intestinal submucosal matrix material isfurther loaded with one or more of chitosan, phenolsulfoethylamine,hemostatic polymer/compound, bioactive molecule, biomimetic activepolypeptide molecule, polyamino acid, polydopamine,polyglycolide-trimethylene carbonate, polyglycolide and copolymersthereof, polycarbopron, poly L-lactide-caprolactone, polyglycolic acid,PPDO and polydioxanone (PDO).

Further, both sides of the antibacterial gel layer are combined with theacellular small intestinal submucosal matrix material and the absorbablefiber layer by hydrogen bonding;

the antibacterial gel layer comprises one or more of gelatin,polypeptide, protein, polyhistidine, fibrin and sodium hyaluronate, thethickness of the antibacterial gel layer is 0.001 mm˜0.01 mm, andsulfadiazine silver is dispersed in the antibacterial gel layer, thecontent of sulfadiazine silver in the antibacterial gel layer is 2˜5 wt.%, and the content of water in the antibacterial gel layer is 1˜5%.

Another aspect of the present invention provides a method for preparingan antibacterial medical biomaterial, comprising:

a) placing the acellular small intestinal submucosal matrix material ina sodium hydroxide solution containing sulfadiazine, and controlling thepH of the solution between 7.1 and 13.0, so as to obtain an intermediatematerial covered with sodium sulfadiazine on the surface and/or insidethereof;

b) after removing the intermediate material obtained in step a from thesolution, washing the same with a sodium chloride solution, and thenperforming a drying treatment, soaking the intermediate material in asilver nitrate solution, and substituting the sodium ions on thesulfadiazine sodium pyrimidine with the silver ions in the silvernitrate to obtain an acellular small intestinal submucosal matrixmaterial loaded with sulfadiazine silver;

c) performing a surface plasma treatment on the acellular smallintestinal submucosal matrix material loaded with sulfadiazine silverobtained in step b to hydroxylate the surface of the acellular smallintestinal submucosal matrix material;

d) coating a layer of antibacterial gel layer on the surface of theacellular small intestinal submucosal matrix material in step c, whereinthe antibacterial gel layer and the acellular small intestinalsubmucosal matrix material are combined by hydrogen bonding;

e) after the absorbable fibers are subjected to plasma treatment,preparing the absorbable fiber solution or gel liquid, and then stirringthe sulfadiazine silver and the absorbable fiber solution or gel liquiduniformly to obtain a mixed solution or a mixed gel liquid;

f) coating the mixed solution or mixed gel liquid in step e on thesurface of the acellular small intestinal submucosal matrix materialwith the antibacterial gel layer prepared in step d, and placing theacellular small intestinal submucosal matrix material in an oven orfreeze-drying to obtain an antibacterial medical biomaterial.

Further, the step b further comprises: soaking the acellular smallintestinal submucosal matrix material loaded with sulfadiazine silver ina compound buffer solution of PH 7.5˜11.0. After the reaction isperformed under the conditions of microwave irradiation, the surface ofthe acellular small intestinal submucosal matrix material loaded withsulfadiazine silver is formed with a compound film, washing and dryingthe submucosal matrix material of the small intestine mucosa loaded withsulfadiazine silver;

the compound buffer solution includes a buffer solution of one or moreof chitosan, phenol sulfoethylamine, hemostatic polymer/compound,bioactive molecule, biomimetic active polypeptide molecule, polyaminoacid, polydopamine, polyglycolide-trimethylene carbonate, polyglycolideand copolymers thereof, polycarbamone, poly L-lactide-caprolactone,polyglycolic acid, PPDO and polydioxanone (PDO).

Further, the reaction in steps a and b is performed in a thermostaticultrasonic washer oscillating in a washing tank, and the ultrasonicshaking condition is that: 25˜35° C. thermostatic reaction for 10˜60minutes, the oscillation frequency is 150˜230 rpm, and the ultrasonicfrequency is 40˜50 kHz.

Further, the molar ratio of sulfadiazine to sodium hydroxide is 1:2˜5,and the molar ratio of sulfadiazine to silver nitrate is 1:2˜5.

Further, the method for preparing the acellular small intestinalsubmucosal matrix material comprises:

a1) immersing a small intestinal submucosal tissue material on an animalbody into physiological saline for washing, placing a clean smallintestinal submucosal tissue material into a closed container, andperforming lyophilization sealing and sterilization storing on the smallintestinal submucosal tissue material in liquid nitrogen;

b1) immersing the defrosted small intestinal submucosal tissue materialinto the decellularization solution for decellularization;

c1) ultrasonically cleaning the small intestinal submucosal tissuematerial after decellularization treatment in a sodium chloridesolution, and stopping cleaning the small intestinal submucosal tissuematerial when the conductivity of the small intestinal submucosal tissuematerial is 1.5 um/s or less to obtain the small intestinal submucosalmatrix material.

Preferably, when decellularization is performed by immersing thedefrosted small intestinal submucosal tissue material into thedecellularization solution, the decellularization is performed bytreating the small intestinal submucosal tissue material obtained instep a1 with the decellularization solution in an environment containingtwo ultrasonic frequency wherein the decellularization solutioncomprises trypsin and PBS solution.

Wherein the mass percentage concentration of trypsin in thedecellularization solution is 0.05˜0.3%, the concentration of PBSsolution is 0.5˜1.5 mmol/L, and the pH value of the decellularizationsolution is 7.0˜8.0.

Further, the decellularization process is performed in a dual-frequencyultrasonic apparatus, in which the low frequency range is 30˜50 KHz, andthe low frequency treatment for 10˜45 min. the high frequency is 70˜100KHz, the high frequency treatment for 15˜45 min, and the temperaturerange of the decellularization solution is 20˜35° C.; ultrasonic poweris 5000 W or more.

Further, the concentration of the sodium chloride solution is 0.01˜0.05mol/L, when the decellularized small intestinal submucosal tissuematerial is ultrasonically cleaned with sodium chloride solution.

The present invention provides an antibacterial medical biomaterial anda method for preparing the same. By loading the sulfadiazine silver onthe acellular small intestinal submucosal matrix material, the acellularsmall intestinal submucosal matrix material loaded with sulfadiazinesilver also has antimicrobial activity consistent with sulfadiazinesilver. The antibacterial gel layer and the absorbable fiber layer arecoated on the outer surface of the acellular small intestinal submucosalmatrix material, so that the moisture-retaining function,liquid-absorbing function, hemostatic function, tear resistance andflexibility of the biomedical material of the biomaterial are improved.Sulfadiazine silver is attached to the absorbable fiber layer, so thatthe medical biomaterial provided by the present invention can be used asan external medicine for treating wound infections relayed by burns orwounds, and can also be used for reducing the incidence of infection byusing a conventional central venous catheter with a sulfadiazine silverantibacterial coating. Therefore, the acellular small intestinalsubmucosal matrix material loaded with sulfadiazine silver also hasantimicrobial activity consistent with sulfadiazine silver, thereby, themedical biomaterial provided by the present invention has antimicrobialand bacteriostatic capability.

Other features and advantages of the invention will be set forth in thedescription which follows and, in part, will be apparent from thedescription, or will be appreciated by practice of the invention.Objects and other advantages of the present invention may be achievedand obtained by structures particularly pointed out in the writtendescription, claims, and drawings.

The technical solutions of the present invention will be furtherdescribed in detail below with reference to the accompanying drawingsand embodiments.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a schematic cross-sectional view of an antibacterial medicalbiomaterial according to an exemplary embodiment of the presentinvention;

FIG. 2 is a schematic top view of an antibacterial medical biomaterialaccording to an exemplary embodiment of the present invention.

The figures are merely illustrative of the invention to facilitateunderstanding of the invention and are not intended to be limiting ofthe invention.

DETAILED DESCRIPTION

Hereinafter, specific embodiments of the present invention will bedescribed in detail with reference to the accompanying drawings, but itshould be understood that the scope of protection of the presentinvention is not limited by the specific embodiments.

Unless otherwise expressly indicated otherwise, throughout thedescription and claims, the term “comprise” or variations thereof suchas “comprising” or “including” and the like will be understood toinclude the stated elements or components, but other elements or othercomponents are not excluded.

Referring to FIGS. 1 and 2 , an aspect of the present invention providesan antibacterial medical biomaterial comprising an acellular smallintestinal submucosal matrix material 1, an antibacterial gel layer 2located on the surface of the acellular small intestinal submucosalmatrix material 1, and an absorbable fiber layer 3 located on thesurface of the antibacterial gel layer 2, sulfadiazine silver isdispersed on the surface of the acellular small intestinal submucosalmatrix material 1 and/or within the acellular small intestinalsubmucosal matrix material 1; the absorbable fiber layer 3 comprisesabsorbable fibers to which sulfadiazine silver is attached, wherein thecontent of sulfadiazine silver in the absorbable fibers 3 is 1 wt. %˜2wt. %.

The absorbable fiber layer 3 has antibacterial properties by attachingsulfadiazine silver to the absorbable fiber, and the arrangement of theabsorbable fiber layer 3 also improves the moisture-retaining function,liquid-absorbing function, hemostatic function, tear resistance andflexibility of the biomedical material. Since the sulfadiazine silverhas good antimicrobial activity against most gram positive bacteria andnegative bacteria, in addition to being widely used as an externalmedicine to treat wound infections secondary to burns or scalds,traditional central venous catheters can also be coated withsulfadiazine silver to reduce the incidence of infections, the acellularsmall intestinal submucosal matrix material loaded with sulfadiazinesilver also has antimicrobial activity consistent with sulfadiazinesilver, thereby, the medical biomaterial provided by the presentinvention has antimicrobial and bacteriostatic capability.

As a preferred embodiment, the absorbable fiber layer 3 has a thicknessof 0.005 mm˜0.05 mm;

Preferably, the absorbable fiber is one or more of hydroxyethylcellulose, oxidized cellulose, and carboxymethyl fiber.

As a preferred embodiment, the content of sulfadiazine silver in theacellular small intestinal submucosal matrix material 1 is 2˜5 wt. %.

In the present invention, the sulfadiazine silver is dispersed on thesurface of the acellular small intestinal submucosal matrix materialland/or within the acellular small intestinal submucosal matrix material1, not only the capacity of loading of sulfadiazine silver is increased,but the durability of loading of sulfadiazine silver is also improved,so that the antibacterial medical biomaterial provided by the presentinvention maintains good antimicrobial performance for a long time.

As a preferred embodiment, the acellular small intestinal submucosalmatrix material 1 is further loaded with one or more of chitosan,phenolsulfoethylamine, hemostatic polymer/compound, bioactive molecule,biomimetic active polypeptide molecule, polyamino acid, polydopamine,polyglycolide-trimethylene carbonate, polyglycolide and copolymersthereof, polycarbamone, poly L-lactide-caprolactone, polyglycolic acid,PPDO, and polydioxanone (PDO). In the present embodiment, the acellularsmall intestinal submucosal matrix material loaded with the abovecompound has various physiological functions such as anti-inflammatory,bacteriostatic, hemostatic, anticancer, pro-growth, and enhancedimmunity in addition to antibacterial properties.

As a preferred embodiment, both sides of the antibacterial gel layer 2are combined with the acellular small intestinal submucosal matrixmaterial 1 and the absorbable fiber layer through hydrogen bonding.

The antibacterial gel layer 2 comprises one or more of gelatin,polypeptide, protein, polyhistidine, fibrin and sodium hyaluronate, thethickness of the antibacterial gel layer 2 is 0.001 mm˜0.01 mm, andsulfadiazine silver is dispersed in the antibacterial gel layer 2, thecontent of sulfadiazine silver in the antibacterial gel layer 2 is 2˜5wt. %, and the content of water in the antibacterial gel layer is 1%˜5%.

Another aspect of the present invention provides a method for preparingan antibacterial medical biomaterial, comprising:

a) placing the acellular small intestinal submucosal matrix material ina sodium hydroxide solution containing sulfadiazine, and controlling thepH of the solution between 7.1 and 13.0, so as to obtain an intermediatematerial loaded with sulfadiazine sodium on the surface and/or insidethereof;

b) after removing the intermediate material obtained in step a from thesolution, washing the same with a sodium chloride solution, and thenperforming a drying treatment, soaking the intermediate material in asilver nitrate solution, and substituting the sodium ions on thesulfadiazine sodium with the silver ions in the silver nitrate to obtainan acellular small intestinal submucosal matrix material loaded withsulfadiazine silver;

c) performing a surface plasma treatment on the acellular smallintestinal submucosal matrix material loaded with sulfadiazine silverobtained in step b to hydroxylate the surface of the acellular smallintestinal submucosal matrix material;

d) coating a layer of antibacterial gel layer on the surface of theacellular small intestinal submucosal matrix material in step c, whereinthe antibacterial gel layer and the acellular small intestinalsubmucosal matrix material are combined by hydrogen bonding;

e) after the absorbable fibers are subjected to plasma treatment,preparing the absorbable fiber solution or gel liquid, and then stirringthe sulfadiazine silver and the absorbable fiber solution or gel liquiduniformly to obtain a mixed solution or a mixed gel liquid;

f) coating the mixed solution or mixed gel liquid in step e on thesurface of the acellular small intestinal submucosal matrix materialwith the antibacterial gel layer prepared in step d, and placing theacellular small intestinal submucosal matrix material in an oven orfreeze-drying to obtain an antibacterial medical biomaterial.

In the present invention, the acellular small intestinal mucosa isplaced in a sodium hydroxide solution containing sulfadiazine, and thesurface of the acellular small intestinal submucosal matrix materialimmersed in the solution is uniformly and firmly loaded withsulfadiazine sodium, so that the surface and/or the interior of theacellular small intestinal submucosal matrix material are coated withsulfadiazine sodium; then, the acellular small intestinal submucosalmatrix material loaded with sulfadiazine sodium is placed in a silvernitrate solution, and the sodium ions on the sulfadiazine sodium arereplaced by silver ions to generate sulfadiazine silver, the sodium ionsand the nitrate form soluble sodium nitrate, the soluble sodium nitratecan be removed, and finally the sulfadiazine silver is synthesized insitu on the acellular small intestinal submucosal matrix material. Inthe method provided by the present invention, the sulfadiazine is notdissolved with the traditional aqueous ammonia, which has the followingadvantages: firstly, the corrosion effect of the aqueous ammonia on theacellular small intestinal submucosal matrix material can be avoided;second, stimulation and corrosion of ammonia water to mucous membranessuch as eyes and nasal cavities of workers can be avoided; thirdly, anenvironmental problem caused by stimulating odor fumes evolving withhigh concentrations of ammonia can be avoided, and in this method, theloading capacity of sulfadiazine silver can be increased without usingcatalyst triflic acid or triflic acid salt, and the safety performancecan be further improved.

As a preferred embodiment, the step b further comprises: soaking theacellular small intestinal submucosal matrix material loaded withsulfadiazine silver in a compound buffer solution of PH 7.5˜11.0, afterthe reaction is performed under the conditions of microwave irradiation,the surface of the acellular small intestinal submucosal matrix materialloaded with sulfadiazine silver is formed with a compound film, washingand drying the acellular small intestinal submucosal matrix materialloaded with sulfadiazine silver;

wherein, the compound buffer solution includes a buffer solution of oneor more of chitosan, phenolsulfoethylamine, hemostatic polymer/compound,bioactive molecule, biomimetic active polypeptide molecule, polyaminoacid, polydopamine, polyglycolide-trimethylene carbonate, polyglycolideand copolymers thereof, polycarbamone, poly L-lactide-caprolactone,polyglycolic acid, PPDO and polydioxanone (PDO).

Upon microwave irradiation, the covalent bonding by the Michael additionreaction or the Schiff base reaction and the non-covalent bondinginclude a combination of van der Waals force and hydrogen bonding, sothat the surface of the acellular small intestinal submucosal matrixmaterial loaded with sulfadiazine silver soaked in the reaction liquidare uniformly and firmly loaded with compounds, thereby, the medicalbiomaterial prepared by the present invention also has variousphysiological functions such as anti-inflammatory, bacteriostatic,hemostatic, anticancer, pro-growth, bacteriostatic, and enhancedimmunity in addition to antibacterial function.

As a preferred embodiment, the reactions in steps a and b are allperformed in a thermostatic ultrasonic washer oscillating in a washingtank, and the ultrasonic shaking condition is that: 25˜35° C.thermostatic reaction for 10˜60 minutes, the oscillation frequency is150˜230 rpm, and the ultrasonic frequency is 40˜50 kHz. In the presentembodiment, the reaction in steps a and c is performed in a thermostaticultrasonic washer capable of oscillating the cleaning tank. The effectof the synthesis reaction can be accelerated, especially whensulfadiazine silver is synthesized in situ, ultrasonic shaking canincrease the probability of silver ions of silver nitrate to contact,react, and bond with sulfadiazine already firmly loaded on the acellularsmall intestinal submucosal matrix material by thousands. The synthesisof sulfadiazine silver is greatly promoted, and the time forsynthesizing sulfadiazine silver in situ on the submucosa material ofthe acellular small intestine mucosa is shortened, and simplifying thestep of synthesizing sulfadiazine silver in situ on the acellular smallintestinal submucosal matrix material. Due to the use of ultrasonicshaking, no physical stirring and external warming are required in thereaction, thereby achieving molecular level stirring and high reactionyield.

As a preferred embodiment, the molar ratio of sulfadiazine to sodiumhydroxide is 1:2˜5, and the molar ratio of sulfadiazine to silvernitrate is 1:2˜5.

As a preferred embodiment, the method for preparing the acellular smallintestinal submucosal matrix material comprises:

a1) immersing a small intestinal submucosal tissue material on an animalbody into physiological saline for washing, placing a clean smallintestinal submucosal tissue material into a closed container, andperforming lyophilization sealing and sterilization storing on the smallintestinal submucosal tissue material in liquid nitrogen;

b1) immersing the defrosted small intestinal submucosal tissue materialinto the decellularization solution for decellularization;

c1) ultrasonically cleaning the small intestinal submucosal tissuematerial after decellularization treatment in a sodium chloridesolution, and stopping cleaning the small intestinal submucosal tissuematerial when the conductivity of the small intestinal submucosal tissuematerial is 1.5 um/s or less to obtain the acellular small intestinalsubmucosal matrix material.

According to the present invention, the small intestine intercepted onthe animal body is immersed in the 4° C. physiological saline forcleaning, preferably, the small intestine is removed and immediatelyafter the small intestine is removed for cleaning, and the animal is notparticularly limited, preferably pig, cattle, sheep, murine or horse.

As a preferable embodiment, when decellularization treatment isperformed by immersing the defrosted small intestinal submucosal tissuematerial into the decellularization solution, the decellularization isperformed by treating the small intestinal submucosal tissue materialobtained in step a1 with the decellularization solution in anenvironment containing two ultrasonic frequency wherein thedecellularization solution comprises trypsin and PBS solution.

As a preferred embodiment, the mass percentage concentration of trypsinin the decellularization solution is 0.05˜0.3%, the concentration of PBSsolution is 0.5˜1.5 mmol/L, and the pH value of the decellularizationsolution is 7.0˜8.0.

As a preferred embodiment, the decellularization process is performed ina dual-frequency ultrasonic apparatus, in which the low frequency rangeis 30˜50 KHz, and the low frequency treatment for 10˜45 min; the highfrequency is 70˜100 KHz, the high frequency treatment for 15˜45 min, andthe temperature range of the decellularization solution is 20˜35° C.;ultrasonic power is 5000 W or more.

As a preferred embodiment, the concentration of the sodium chloridesolution is 0.01˜0.05 mol/L when the decellularized small intestinalsubmucosal tissue material is ultrasonically cleaned with sodiumchloride solution.

As a preferred embodiment, the lyophilization temperature in step a1 is−75˜−80° C., and the lyophilization time is 20˜30 h.

As a preferred embodiment, the sterilization is to sterilize the sealedsmall intestinal submucosal tissue material through linear accelerationwith a radiation dose of 8000 rads.

Hereinafter, an antibacterial medical biomaterial provided by thepresent invention will be described in detail by way of examples.

Example 1

A method for preparing an antibacterial medical biomaterial, comprising:

(1) immersing a small intestinal submucosal tissue material on a porcinebody into physiological saline for washing, placing a clean smallintestinal submucosal tissue material into a closed container, andperforming lyophilization sealing and sterilization on the smallintestinal submucosal tissue material in liquid nitrogen, wherein thelyophilization temperature is −75° C., the lyophilization time is 25 h,and the sterilization is to sterilize the sealed small intestinalsubmucosal tissue material through linear acceleration with a radiationdose of 8000 rads.

(2) treating the small intestinal submucosal tissue material obtained instep a1 with decellularization in an environment containing twoultrasonic frequencies with different frequencies for decellularization;wherein the decellularization solution comprises trypsin and PBSsolution, the mass percentage concentration of trypsin in thedecellularization solution is 0.05%, the concentration of PBS solutionis 1 mmol/L, and the pH value of the decellularization solution is 7.0.

(3) ultrasonically cleaning the decellularized small intestinalsubmucosal tissue material in sodium chloride solution, theconcentration of sodium chloride solution is 0.03 mol/L, and when theelectrical conductivity of the small intestinal submucosal tissuematerial is 1.5 um/s or less, stopping cleaning the small intestinalsubmucosal tissue material to obtain the acellular small intestinalsubmucosal matrix material.

Wherein the decellularization process is performed in a dual-frequencyultrasonic apparatus, in which the low frequency range is 30 KHz and thelow frequency treatment for 25 min, the high frequency is 90 KHz, thehigh frequency treatment for 15 min, and the temperature range of thedecellularization solution is 30° C.; ultrasonic power is 5000 W ormore.

(4) after placing the acellular small intestinal submucosal matrixmaterial in a sodium hydroxide solution containing sulfadiazine, placingin a thermostatic ultrasonic washer capable of oscillating the cleaningtank, the pH of the solution was controlled to 7.1, and the temperaturewas controlled to 25° C. The reaction was performed for 10 minutes, theoscillation frequency was 150 rpm, and the ultrasonic frequency was 40kHz. obtaining a surface and/or an intermediate material coated withsulfadiazine sodium, the molar ratio of sulfadiazine to sodium hydroxideis 1:2, the molar ratio of sulfadiazine to silver nitrate is 1:2.

(5) after the intermediate material is removed from the solution, theintermediate material is washed with a sodium chloride solution. Afterthe drying treatment, the intermediate material is immersed in a silvernitrate solution, and placed in a thermostatic ultrasonic washer capableof oscillating the cleaning tank, the temperature was controlled to 25°C., the reaction was performed for 10 minutes, and the oscillationfrequency was 150 rpm. The ultrasonic frequency is 40 KHz, and thesodium ions on the sulfadiazine sodium molecule are replaced by silverions in silver nitrate. An acellular small intestinal submucosal matrixmaterial loaded with sulfadiazine silver was obtained.

(6) performing a surface plasma treatment on the acellular smallintestinal submucosal matrix material loaded with sulfadiazine silver,and so as to hydroxylate the surface of the acellular small intestinalsubmucosal matrix material.

(7) coating a layer of antibacterial gel layer on the surface of theacellular small intestinal submucosal matrix material, wherein theantibacterial gel layer and the acellular small intestinal submucosalmatrix material are combined by hydrogen bonding.

(8) after the absorbable fibers are subjected to plasma treatment,preparing the absorbable fiber solution or gel liquid, and then stirringthe sulfadiazine silver and the absorbable fiber solution or gel liquiduniformly to obtain a mixed solution or a mixed gel liquid.

(9) coating a mixed solution or a mixed gel liquid on the surface of theacellular small intestinal submucosal matrix material with theantibacterial gel layer, and placing the acellular small intestinalsubmucosal matrix material in an oven or freeze-drying to obtain anantibacterial medical biomaterial.

Wherein, the step 5 further comprises: soaking the acellular smallintestinal submucosal matrix material loaded with sulfadiazine silver ina compound buffer solution of PH 7.5, performing a reaction undermicrowave irradiation conditions, and after the surface of the submucousmatrix material of the acellular small intestine mucous membranesupported with sulfadiazine silver is formed with a compound film, theacellular small intestinal submucosal matrix material loaded withsulfadiazine silver is taken out, washed and dried;

The compound buffer solution includes a buffer solution of one or moreof chitosan, phenol sulfoethylamine, hemostatic polymer/compound,bioactive molecule, biomimetic active polypeptide molecule, polyaminoacid, polydopamine, polyglycolide-trimethylene carbonate, polyglycolideand copolymers thereof, polycarbamone, poly L-lactide-caprolactone,polyglycolic acid, PPDO and polydioxanone (PDO).

Example 2

A method for preparing an antibacterial medical biomaterial, comprising:

(1) immersing a small intestinal submucosal tissue material on a porcinebody into physiological saline for washing, placing a clean smallintestinal submucosal tissue material into a closed container, andperforming lyophilization sealing and sterilization on the smallintestinal submucosal tissue material in liquid nitrogen, wherein thelyophilization temperature is −78° C., the lyophilization time is 25 h,and the sterilization is to sterilize the sealed small intestinalsubmucosal tissue material through linear acceleration with a radiationdose of 8000 rads.

(2) treating the small intestinal submucosal tissue material obtained instep a1 with decellularization in an environment containing twoultrasonic frequencies with different frequencies for decellularization;wherein the decellularization solution comprises trypsin and PBSsolution, the mass percentage concentration of trypsin in thedecellularization solution is 0.2%, the concentration of PBS solution is0.5 mmol/L, and the pH value of the decellularization solution is 7.0.

(3) ultrasonically cleaning the decellularized small intestinalsubmucosal tissue material in sodium chloride solution, theconcentration of sodium chloride solution is 0.01 mol/L, and when theelectrical conductivity of the small intestinal submucosal tissuematerial is 1.5 um/s or less, stopping cleaning the small intestinalsubmucosal tissue material to obtain the acellular small intestinalsubmucosal matrix material.

Wherein the decellularization process is performed in a dual-frequencyultrasonic apparatus, in which the low frequency range is 40 KHz and thelow frequency treatment is 10 min. the high frequency is 70 KHz, thehigh frequency treatment is 35 min, and the temperature range of thedecellularized liquid is 20° C. centigrade; Ultrasonic power is 5000 Wor more.

(4) after placing the acellular small intestinal submucosal matrixmaterial in a sodium hydroxide solution containing sulfadiazine, placingin a thermostatic ultrasonic washer capable of oscillating the cleaningtank, the pH of the solution was controlled to 11, the temperature wascontrolled to 30° C., and the reaction was performed for 30 minutes. Theoscillation frequency is 200 rpm, and the ultrasonic frequency is 45KHz, so as to obtain an intermediate material having a surface and/or aninner coated with sulfadiazine sodium. The molar ratio of sulfadiazineto sodium hydroxide is 1:3, the molar ratio of sulfadiazine to silvernitrate was 1:4.

(5) after the intermediate material is removed from the solution, theintermediate material is washed with a sodium chloride solution. Afterthe drying treatment, the intermediate material is immersed in a silvernitrate solution, and placed in a thermostatic ultrasonic washer capableof oscillating the cleaning tank, the temperature was controlled to 30°C., the reaction was performed for 30 minutes, and the oscillationfrequency was 200 rpm. The ultrasonic frequency is 45 KHZ, and thesodium ions on the sulfadiazine sodium molecule are replaced by silverions in silver nitrate. An acellular small intestinal submucosal matrixmaterial loaded with sulfadiazine silver was obtained.

(6) performing a surface plasma treatment on the acellular smallintestinal submucosal matrix material loaded with sulfadiazine silver,and so as to hydroxylate the surface of the acellular small intestinalsubmucosal matrix material;

(7) coating a layer of antibacterial gel layer on the surface of theacellular small intestinal submucosal matrix material, wherein theantibacterial gel layer and the acellular small intestinal submucosalmatrix material are combined by hydrogen bonding;

(8) after the absorbable fibers are subjected to plasma treatment,preparing the absorbable fiber solution or gel liquid, and then stirringthe sulfadiazine silver and the absorbable fiber solution or gel liquiduniformly to obtain a mixed solution or a mixed gel liquid;

(9) coating a mixed solution or a mixed gel liquid on the surface of theacellular small intestinal submucosal matrix material with theantibacterial gel layer on the prepared surface, and placing theacellular small intestinal submucosal matrix material in an oven orfreeze-drying to obtain an antibacterial medical biomaterial.

Wherein, the step 5 further comprises: soaking the acellular smallintestinal submucosal matrix material loaded with sulfadiazine silver ina compound buffer solution having a pH of 9, performing a reaction undermicrowave irradiation conditions, and after the surface of the submucousmatrix material of the acellular small intestine mucous membranesupported with sulfadiazine silver is formed with a compound film, theacellular small intestinal submucosal matrix material loaded withsulfadiazine silver is taken out, washed and dried;

The compound buffer solution includes a buffer solution of one or moreof chitosan, phenol sulfoethylamine, hemostatic polymer/compound,bioactive molecule, biomimetic active polypeptide molecule, polyaminoacid, polydopamine, polyglycolide-trimethylene carbonate, polyglycolideand copolymers thereof, polycarbamone, polyL-lactide-caprolactone,polyglycolic acid, PPDO and polydioxanone (PDO).

Example 3

A method for preparing an antibacterial medical biomaterial, comprising:

(1) immersing a small intestinal submucosal tissue material on a porcinebody into physiological saline for washing, placing a clean smallintestinal submucosal tissue material into a closed container, andperforming lyophilization sealing and sterilization on the smallintestinal submucosal tissue material in liquid nitrogen, wherein thelyophilization temperature is −80° C., the sterilization is to sterilizethe sealed small intestinal submucosal tissue material through linearacceleration with a radiation dose of 8000 rads.

(2) treating the small intestinal submucosal tissue material obtained instep a1 with decellularization in an environment containing twoultrasonic frequencies with different frequencies for decellularization;wherein the decellularization solution comprises trypsin and PBSsolution, the mass percentage concentration of trypsin in thedecellularization solution is 0.3%, the concentration of PBS solution is1.5 mmol/L, and the pH value of the decellularization solution is 8.0.

(3) ultrasonically cleaning the decellularized small intestinalsubmucosal tissue material in sodium chloride solution, theconcentration of sodium chloride solution is 0.05 mol/L, and when theelectrical conductivity of the small intestinal submucosal tissuematerial is 1.5 um/s or less, stopping cleaning the small intestinalsubmucosal tissue material to obtain the acellular small intestinalsubmucosal matrix material.

Wherein the decellularization process is performed in a dual-frequencyultrasonic apparatus, in which the low frequency range is 50 KHz and thelow frequency treatment for 45 min; the high frequency is 100 KHz, thehigh frequency treatment for 45 min, and the temperature range of thedecellularization solution is 35° C.; ultrasonic power is 5000 W ormore.

(4) after placing the acellular small intestinal submucosal matrixmaterial in a sodium hydroxide solution containing sulfadiazine, placingin a thermostatic ultrasonic washer capable of oscillating the cleaningtank, the pH of the solution was controlled to 13.0, the temperature wascontrolled to 35° C. centigrade, and the reaction was performed for 60minutes, the oscillation frequency is 230 rpm, and the ultrasonicfrequency is 50 KHz, so as to obtain an intermediate material having asurface and/or an inner coated with sulfadiazine sodium. The molar ratioof sulfadiazine to sodium hydroxide is 1:5. The molar ratio ofsulfadiazine to silver nitrate was 1:5.

(5) after the intermediate material is removed from the solution, theintermediate material is washed with a sodium chloride solution. Afterthe drying treatment, the intermediate material is immersed in a silvernitrate solution, and placed in a thermostatic ultrasonic washer capableof oscillating the cleaning tank, the temperature was controlled to 35°C. centigrade, the reaction was performed for 60 minutes, and theoscillation frequency was 230 rpm. The ultrasonic frequency is 50 KHZ,and the sodium ions on the sulfadiazine sodium molecule are replaced bysilver ions in silver nitrate. An acellular small intestinal submucosalmatrix material loaded with sulfadiazine silver was obtained.

(6) performing a surface plasma treatment on the acellular smallintestinal submucosal matrix material loaded with sulfadiazine silver,and so as to hydroxylate the surface of the acellular small intestinalsubmucosal matrix material.

(7) coating a layer of antibacterial gel layer on the surface of theacellular small intestinal submucosal matrix material, wherein theantibacterial gel layer and the acellular small intestinal submucosalmatrix material are combined by hydrogen bonding.

(8) after the absorbable fibers are subjected to plasma treatment,preparing the absorbable fiber solution or gel liquid, and then stirringthe sulfadiazine silver and the absorbable fiber solution or gel liquiduniformly to obtain a mixed solution or a mixed gel liquid.

(9) coating a mixed solution or a mixed gel liquid on the surface of theacellular small intestinal submucosal matrix material with theantibacterial gel layer on the prepared surface, and placing theacellular small intestinal submucosal matrix material in an oven orfreeze-drying to obtain an antibacterial medical biomaterial.

Wherein, the step 5 further comprises: soaking the acellular smallintestinal submucosal matrix material loaded with sulfadiazine silver ina compound buffer solution having a pH of 11.0, performing a reactionunder microwave irradiation conditions, and after the surface of thesubmucous matrix material of the acellular small intestine mucousmembrane supported with sulfadiazine silver is formed with a compoundfilm, the acellular small intestinal submucosal matrix material loadedwith sulfadiazine silver is taken out, washed and dried;

The compound buffer solution includes a buffer solution of one or moreof chitosan, phenol sulfoethylamine, hemostatic polymer/compound,bioactive molecule, biomimetic active polypeptide molecule, polyaminoacid, polydopamine, polyglycolide-trimethylene carbonate, polyglycolideand copolymers thereof, polycarbamone, poly L-lactide-caprolactone,polyglycolic acid, PPDO and polydioxanone (PDO).

Product Antimicrobial Testing

The antibacterial activity of the antibacterial medical biomaterialprepared in the examples 1˜3 is tested, and the test results are shownin table 1:

TABLE 1 results of antibacterial tests on the products of the examplesExample 1 Example 2 Example 3 Gram-positive 96.9 96.3 97.1 bacteriaNegative bacteria 97.8 98.1 97.9

The antibacterial property of the antibacterial medical biomaterial togram-positive bacteria and gram-negative bacteria can still reach morethan 95 percent, which shows that the medical biomaterial provided bythe invention has good antibacterial effect.

Finally, it should be noted that the above embodiments are only forillustrating the technical solutions of the present invention and notfor limiting, and although the present invention is described in detailwith reference to examples, it should be understood by those skilled inthe art that modifications or equivalent substitutions may be made tothe technical solutions of the present invention without departing fromthe spirit and scope of the technical solutions of the presentinvention, which should be covered by the claims of the presentinvention.

The invention claimed is:
 1. A method for preparing an antibacterialmedical biomaterial, comprising: a) placing an acellular smallintestinal submucosal matrix material in a sodium hydroxide solutioncontaining sulfadiazine, and controlling pH of the solution between 7.1and 13.0, so as to obtain an intermediate material loaded withsulfadiazine sodium on the surface and/or inside thereof; b) afterremoving the intermediate material obtained in step a from the solution,washing the same with a sodium chloride solution, and then performing adrying treatment, soaking the intermediate material in a silver nitratesolution, and substituting sodium ions on a sulfadiazine sodiumpyrimidine with silver ions in the silver nitrate to obtain an acellularsmall intestinal submucosal matrix material loaded with sulfadiazinesilver; c) performing a surface plasma treatment on the acellular smallintestinal submucosal matrix material loaded with sulfadiazine silverobtained in step b to hydroxylate a surface of the acellular smallintestinal submucosal matrix material; d) coating a layer ofantibacterial gel layer on the surface of the acellular small intestinalsubmucosal matrix material in step c, wherein the antibacterial gellayer and the acellular small intestinal submucosal matrix material arecombined by hydrogen bonding; e) after absorbable fibers are subjectedto plasma treatment, preparing the absorbable fiber solution or gelliquid, and then stirring the sulfadiazine silver and the absorbablefiber solution or gel liquid uniformly to obtain a mixed solution or amixed gel liquid; f) coating the mixed solution or mixed gel liquid instep e on the surface of the acellular small intestinal submucosalmatrix material with the antibacterial gel layer prepared in step d, andplacing the acellular small intestinal submucosal matrix material in anoven or freeze-drying to obtain an antibacterial medical biomaterial. 2.The method for preparing an antibacterial medical biomaterial accordingto claim 1, wherein the step b further comprises: soaking the acellularsmall intestinal submucosal matrix material loaded with sulfadiazinesilver in a compound buffer solution of PH 7.5˜11.0; after the reactionis performed under the conditions of microwave irradiation, the surfaceof the acellular small intestinal submucosal matrix material loaded withsulfadiazine silver is formed with a compound film, washing and dryingthe acellular small intestinal submucosal matrix material loaded withsulfadiazine silver; the compound buffer solution includes a buffersolution of one or more of chitosan, phenol sulfoethylamine, hemostaticpolymer/compound, bioactive molecule, biomimetic active polypeptidemolecule, polyamino acid, polydopamine, polyglycolide-trimethylenecarbonate, polyglycolide and copolymers thereof, polycarbamone, polyL-lactide-caprolactone, polyglycolic acid, PPDO and PDO.
 3. The methodfor preparing an antibacterial medical biomaterial according to claim 1,wherein the reactions in steps a and b are all performed in athermostatic ultrasonic washer oscillating in a washing tank, and theultrasonic shaking condition is that: 25˜35° C. thermostatic reactionfor 10˜60 minutes, the oscillation frequency is 150˜230 rpm, and theultrasonic frequency is 40˜50 kHz.
 4. The method for preparing anantibacterial medical biomaterial according to claim 1, wherein themolar ratio of sulfadiazine to sodium hydroxide is 1:2˜5, and the molarratio of sulfadiazine to silver nitrate is 1:2˜5.
 5. The method forpreparing an antibacterial medical biomaterial according to claim 1,wherein a method for preparing an acellular small intestinal submucosalmatrix material comprises: a1) immersing a small intestinal submucosaltissue material on an animal body into physiological saline for washing,placing a clean small intestinal submucosal tissue material into aclosed container, and performing lyophilization sealing andsterilization storing on the small intestinal submucosal tissue materialin liquid nitrogen; b1) immersing the defrosted small intestinalsubmucosal tissue material into the decellularization solution fordecellularization; c1) ultrasonically cleaning the small intestinalsubmucosal tissue material after decellularization treatment in a sodiumchloride solution, and stopping cleaning the small intestinal submucosaltissue material when the conductivity of the small intestinal submucosaltissue material is 1.5 um/s or less to obtain the acellular smallintestinal submucosal matrix material; wherein when decellularization isperformed by immersing the defrosted small intestinal submucosal tissuematerial into the decellularization solution, the decellularization isperformed by treating the small intestinal submucosal tissue materialobtained in step a1 with the decellularization solution in anenvironment containing two ultrasonic frequency wherein thedecellularization solution comprises trypsin and PBS solution; whereinthe mass percentage concentration of trypsin in the decellularizationsolution is 0.05˜0.3%, the concentration of PBS solution is 0.5˜1.5mmol/L, and the pH value of the decellularization solution is 7.0˜8.0.6. The method for preparing an antibacterial medical biological materialaccording to claim 5, wherein the decellularization process is performedin a dual-frequency ultrasonic apparatus, wherein a low frequency rangeis 30˜50 KHz, and a low frequency treatment for 10˜45 min; a highfrequency is 70˜100 KHz, a high frequency treatment for 15˜45 min, and atemperature range of the decellularization solution is 20˜35° C.;ultrasonic power is 5000 W or more.
 7. The method for preparing anantibacterial medical biomaterial according to claim 5, wherein aconcentration of the sodium chloride solution is 0.01˜0.05 mol/L, whenthe decellularized small intestinal submucosal tissue material isultrasonically cleaned with sodium chloride solution.